Identification of cell culture conditions to control protein aggregation of IgG fusion proteins expressed in Chinese hamster ovary cells

Tailoring recombinant protein quality by rational media design

“…Other compounds, such as DMSO (1-8%, v/v) and glycerol (1-2%, v/v), exhibited protein stabilization capabilities 139,173 . In contrast, the addition of copper into the cell culture medium could slightly increase aggregation, whereas a supplementation of the culture medium with cysteine known for to its mild reducing characteristics on disulfide bond bridges, resulted in a substantial decrease in aggregate content and a corresponding increase in single chain species 54 . Cystine, the oxidized form of cysteine, also reduced high-molecularweight forms (HMW), but in contrast to cysteine, induced a much lower single-chain content increase.…”

“…Many authors have published extensive reviews, depicting the current state of the art and strategies, which focus on the cell line 1,7,16,45 and the cell culture parameters 18,51 . Furthermore, through the concentration adjustment of selected media components, and in some cases by supplementing the medium with specific co-factors, it is possible to adjust the glycosylation profile 52 , the charge variants 53 , the aggregation level 46,54 , and the abundance of LMW species 55 . Figure 1.1 outlines the three main strategies allowing to affect cell culture process performance and to tailor the quality attributes of therapeutic molecules.…”

“…Various reducing and oxidizing reagents including glutathionine, cysteine and copper sulfate proved to reduce the formation of protein aggregates or increase their stability in CHO cell culture harvests 54 . Other compounds, such as DMSO (1-8%, v/v) and glycerol (1-2%, v/v), exhibited protein stabilization capabilities 139,173 .…”

“…Combining cystine concentration and culture temperature shift was even more effective in decreasing protein aggregation. Furthermore, it resulted in greater sialylation and higher harvest titer 54 . Iron plays a major role in inhibiting the formation of aggregate forms, as shown in long-term drug substance stability studies, where the abundance of high-molecularweight species was significantly lower at low iron concentrations (0-4 ppm) in the presence of a chelator 174 .…”

“…It is also important to keep in mind that the productivity of recombinant cell culture processes has substantially increased from 50 mg/L in batch to titers of 10 to 13 g/L nowadays in fed-batch culture of 2 to 3 weeks 33,34 . Moreover, the literature describes how cell culture conditions can be identified to control protein aggregation in CHO cell cultures 54 . A variety of chromatographic resins have been designed for efficient aggregate removal.…”